What Is A Knockin?
What Is a Knockin Mouse and Why Is It Used?
Understanding just what is a knockin method and a knockin mouse can help us further our understanding of some of the most advanced research methods used in modern science. As researchers continue to find out more about the human genome and the role that each gene plays in supporting life, transgenic and knockin mouse models continue to play an essential role in helping with that process of discovery. Used in genetic engineering, knockin mice are designed to use specific targeting of knockin gene locations that can be utilized to shed light on the complexity of the human genome through the detailed study of the similar mouse genome and its subsequent genetic mutations resulting from knockin gene insertion.
An Overview of the Gene Knockin Process
One of the first questions we have to ask is what is a knockin and why is it deemed so useful in the world of genetic research and engineering. Knockins are typically used in biology, genetic research and cloning to replace a DNA sequence at a specific locus within the genome, as opposed to the less accurate transgenic methods of replacement. Unlike in knockout animal models, a knockin will use ES cells to infiltrate a genetic mutation often established through a targeted insertion that allows researchers to insert an entirely new gene into a particular locus within the genome. Knockin models are most commonly used to track diseases and their effects on live subjects – usually knockin laboratory mouse models.
How Does the Gene Knockin Method Work?
What is a knockin gene replacement and how is it used? The process of creating a knockin animal – most often a knockin mouse – can be extremely complex when regarded from a practical standpoint, despite the seemingly simple strategy it requires. The technique involves a slight change when compared to the classic knockout method developed by Evans, Capecchi and Smithies. The process involves targeted gene replacement mostly through homologous recombination. Typically, procedures such as Cre recombinase with gene vectors and loxP flanking sites are used, but some methods also use the lox511 sequence, which is equally effective in Cre-Lox recombination.
What Are the Knockin Processes’ Primary Uses?
While we have already looked at a general outline of the role knockins have in studying disease, the question remains, what is a knockin used for most frequently? Knockin gene replacement methods have had a fair degree of success, and researchers are looking forward to using them for a variety of clinical applications. One example is the insertion of the human version of the immunoglobuline gene into mice for the purpose of studying antibodies and their ability to fend off viruses and other infections. Other methods focus on controlling and studying regulatory elements like promotors.
The Diversity of Knockin Mouse Models
Conventional knockins involve a gene being introduced at endogenous locuses. However, there are other methods that are in use and have to be mentioned when it comes to aiming to have a complete picture. Point mutation affects the function of particular proteins by one or more nucleotides. At the same time, reporter models focus more on monitoring specific genetic events. Finally, humanized knockin mouse models can be used to introduce a human gene into the mouse genome. With this remarkable level of diversity, knowing what is a knockin animal, choosing the right type of model and studying the creation of new knockin mouse models could even save people’s lives.